Alvarado-Marchena Luis, Furman Bradley T, Breitbart Mya
College of Marine Science, University of South Florida, St. Petersburg, Florida, USA.
Florida Fish and Wildlife Conservation Commission, Florida Fish and Wildlife Research Institute, St. Petersburg, Florida, USA.
mBio. 2025 Jan 8;16(1):e0282824. doi: 10.1128/mbio.02828-24. Epub 2024 Dec 11.
Seagrasses are a polyphyletic group of marine flowering plants that play crucial roles in nearshore ecology, yet their interactions with viruses remain largely unexplored. This study presents the construction and characterization of an infectious cDNA clone of the potexvirus turtle grass virus X (TGVX). The complete genome of this positive-sense single-stranded RNA virus was amplified from field samples of and assembled into a pLX-based mini binary vector using a multi-fragment directional cloning strategy, resulting in the infectious clone pLX-TGVX. Agroinfection assays of potexvirus-free plants resulted in systemic infections by TGVX, as confirmed by multiplex RT-PCR experiments and phenotypic changes reflecting virus-induced symptoms. Ultrastructural studies also demonstrated significant cytopathological changes resulting from TGVX infection, including chloroplast swelling, reduced thylakoid grana, and the presence of viral replication organelles and filamentous virus-like particles. The development of the TGVX infectious clone offers a novel tool for investigating the impact of this virus on seagrass health and productivity. This study demonstrates the first successful agroinfection of a marine plant with an infectious clone, creating a new avenue for studying viruses identified through sequence-based surveys and paving the way for exploring the ecological significance of viral infection in these critical marine ecosystems.IMPORTANCEThis study pioneers the construction of an infectious clone of turtle grass virus X and describes its application in the natural marine plant host, . The creation of this infectious clone not only provides a valuable tool for marine plant virology research but also opens new avenues for exploring the influence of viral infections on the health and productivity of seagrass meadows. Given that seagrasses play a crucial role in sediment stabilization, nutrient cycling, and habitat provisioning, understanding the impact of viruses on these ecosystems is essential for their effective conservation and management. This methodological advance enables detailed studies of viral replication, virus-host interactions, and the broader ecological implications of viral infections in marine plants.
海草是一类多源的海洋开花植物，在近岸生态系统中发挥着关键作用，然而它们与病毒的相互作用在很大程度上仍未得到探索。本研究展示了龟草病毒X（TGVX）感染性cDNA克隆的构建及特性分析。这种正义单链RNA病毒的完整基因组从[未提及的植物名称]的田间样本中扩增出来，并使用多片段定向克隆策略组装到基于pLX的微型双元载体中，得到感染性克隆pLX-TGVX。对无马铃薯X病毒的[未提及的植物名称]植株进行的农杆菌介导感染试验导致TGVX引发系统性感染，多重逆转录聚合酶链反应实验以及反映病毒诱导症状的表型变化证实了这一点。超微结构研究还表明，TGVX感染导致了显著的细胞病理学变化，包括叶绿体肿胀、类囊体基粒减少，以及病毒复制细胞器和丝状病毒样颗粒的存在。TGVX感染性克隆的开发为研究这种病毒对海草健康和生产力的影响提供了一种新工具。本研究证明了首次成功地用感染性克隆对海洋植物进行农杆菌介导感染，为研究通过基于序列的调查鉴定出的病毒开辟了一条新途径，并为探索这些关键海洋生态系统中病毒感染的生态意义铺平了道路。
重要性
本研究率先构建了龟草病毒X的感染性克隆，并描述了其在天然海洋植物宿主[未提及的植物名称]中的应用。这种感染性克隆的创建不仅为海洋植物病毒学研究提供了一个有价值的工具，还为探索病毒感染对海草草甸健康和生产力的影响开辟了新途径。鉴于海草在沉积物稳定、养分循环和栖息地提供方面发挥着关键作用，了解病毒对这些生态系统的影响对于它们的有效保护和管理至关重要。这一方法学进展能够对海洋植物中病毒的复制、病毒-宿主相互作用以及病毒感染的更广泛生态影响进行详细研究。
